赖氨酸对奶牛乳腺上皮细胞内乳糖合成相关基因表达的影响

陈璐, 赵艳丽, 郭晓宇, 史彬林, 闫素梅

doi:10.3969/j.issn.1006-267x.2018.09.029

动物营养学报 >

2018 , Vol. 30 >Issue 9: 3593 - 3598

DOI: https://doi.org/10.3969/j.issn.1006-267x.2018.09.029

分子营养 Molecular Nutrition

赖氨酸对奶牛乳腺上皮细胞内乳糖合成相关基因表达的影响

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内蒙古农业大学动物科学学院, 呼和浩特 010018

陈璐(1990-),女,山西襄汾人,硕士研究生,从事奶牛营养研究。E-mail:1510560671@qq.com

收稿日期: 2018-02-14

网络出版日期: 2018-09-20

基金资助

国家奶业"973计划"项目（2011CB1008003）

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Effects of Lysine on Expression of Genes Related to Lactose Synthesis in Bovine Mammary Epithelial Cells

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College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China

Received date: 2018-02-14

Online published: 2018-09-20

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摘要

本试验旨在研究赖氨酸（Lys）对奶牛乳腺上皮细胞（BMECs）内乳糖合成相关基因表达的影响，深入探讨Lys对乳糖合成的影响机理。将第3代BMECs随机分为6个组，每组6个重复，各组细胞培养液中Lys的终浓度分别为0.5（对照）、1.0、2.0、4.0、8.0和16.0 mmol/L，细胞在37℃、5% CO₂条件下培养48 h。结果表明：适宜浓度Lys对乳糖含量和葡萄糖转运载体蛋白1（GLUT1）、已糖激酶Ⅰ（HKⅠ）和已糖激酶Ⅱ（HKⅡ）基因表达量的影响呈显著的剂量依赖关系（P<0.05）；方差分析结果显示，添加Lys显著或趋于显著影响乳糖含量及乳糖合成相关基因表达。与对照组相比，2.0~16.0 mmol/L组乳糖含量较高（0.05 < P < 0.10），4.0~16.0 mmol/L组GLUT1基因表达量显著升高（P<0.05），2.0~8.0 mmol/L组α-乳清白蛋白（LALBA）和β-1，4-半乳糖基转移酶-1（β-4GALT1）基因表达量显著升高（P<0.05），8.0~16.0 mmol/L组HKⅡ基因表达量显著升高（P<0.05）；但1.0~2.0 mmol/L组HKⅡ基因表达量显著下降（P<0.05），4.0~16.0 mmol/L组HKⅠ基因表达量下降，尤以16.0 mmol/L组显著低于对照组（P<0.05）。综上所述，Lys对乳糖含量和GLUT1、HKⅠ和HKⅡ基因表达量的影响存在显著的剂量依赖，Lys浓度为2.0~8.0 mmol/L时，对BMECs内乳糖合成促进效果较好。

关键词： 奶牛; 乳腺上皮细胞; 赖氨酸; 乳糖; 基因表达

本文引用格式

陈璐, 赵艳丽, 郭晓宇, 史彬林, 闫素梅 . 赖氨酸对奶牛乳腺上皮细胞内乳糖合成相关基因表达的影响[J]. 动物营养学报, 2018 , 30(9) : 3593 -3598 . DOI: 10.3969/j.issn.1006-267x.2018.09.029

Abstract

The objective of this study was to determine the effect of lysine (Lys) on the expression of genes related to lactose synthesis in bovine mammary epithelial cells (BMECs), and to discuss the mechanism of the effect of Lys on lactose synthesis. The third generation BMECs were divided into 6 groups with 6 repetitions, and were cultured in the medium at concentrations of 0.5 (control), 1.0, 2.0, 4.0, 8.0 and 16.0 mmol/L Lys, respectively, for 48 h cultivation at 37℃ and 5% CO₂. The results showed as follows:the proper concentration of Lys had a significant dose-dependent effect on the content of lactose and the expression levels of glucose transporter 1 (GLUT1), hexokinase Ⅰ (HKⅠ) and hexokinase Ⅱ (HKⅡ) genes (P<0.05); the variance analysis results showed that increasing Lys concentration had a significant or tend to significant effect on the content of lactose and the expression of genes related to lactose synthesis. Compared with the control, the content of lactose in 2.0 to 16.0 mmol/L Lys groups was higher (0.05 < P < 0.10), the expression level of GLUT1 gene in 4.0 to 16.0 mmol/L Lys groups increased significantly (P<0.05), the expression levels of α-lactalbumin (LALBA) and β-1,4-galactosyltransferase 1 (β-4GALT1) genes in 2.0 to 8.0 mmol/L Lys groups increased significantly (P<0.05), and the expression level of HKⅡ gene in 8.0 to 16.0 mmol/L Lys groups increased significantly (P<0.05); the expression level of HKⅡ gene in 1.0 to 2.0 mmol/L Lys groups was inhibited significantly (P<0.05); the expression of HKⅠ gene in 4.0 to 16.0 mmol/L Lys groups was inhibited, especially for that in 16.0 mmol/L Lys group, which was significantly lower than that in the control (P<0.05). In conclusion, it shows that the Lys significantly improves the content of lactose, GLUT1, HKⅠ and HKⅡ genes expression in dose-dependent manner and the optimal Lys concentrations for lactose synthesis is 2.0 to 8.0 mmol/L.

Key words： dairy cow; bovine mammary epithelial cells; lysine; lactose; gene expression

参考文献

[1] BRODBECK U,DENTON W L,TANAHASHI N,et al.The isolation and identification of the B protein of lactose synthetase as α-lactalbumin[J].Journal of Biological Chemistry,1967,242(7):1391-1397.

[2] VILOTTE J.Lowering the milk lactose content in vivo:potential interests,strategies and physiological consequences[J].Reproduction,Nutrition,Development,2002,42(2):127-132.

[3] WICKRAMASINGHE S,RINCON G,ISLAS-TREJO A,et al.Transcriptional profiling of bovine milk using RNA sequencing[J].BMC Genomics,2012,13:45.

[4] MAXIN G,RULQUIN H,GLASSER F,et al.Response of milk fat concentration and yield to nutrient supply in dairy cows[J].Animal,2011,5(8):1299-1310.

[5] ALLAUDDIN,BABAR M E,BHATTI J A,et al.Effect of methionine supplementation on milk production and composition in nili-ravi buffaloes[J].Pakistan Journal of Zoology Supplementry Series,2009,9:413-416.

[6] 王立娜.氨基酸与STAT5A基因互作对奶牛乳腺上皮细胞泌乳的调节作用及机理[D].博士学位论文.哈尔滨:东北农业大学,2014.

[7] 云伏雨.瘤胃保护赖氨酸对泌乳中期奶牛生产性能和血液生化指标的影响[D].硕士学位论文.呼和浩特:内蒙古农业大学,2011.

[8] SHENG R,YAN S M,QI L Z,et al.Effect of the ratios of acetate and β-hydroxybutyrate on the expression of milk fat-and protein-related genes in bovine mammary epithelial cells[J].Czech Journal of Animal Science,2015,60(12):531-541.

[9] 高海娜.亮氨酸、组氨酸、赖氨酸和蛋氨酸对奶牛乳腺上皮细胞中酪蛋白合成的影响及调控机理研究[D].硕士学位论文.兰州:甘肃农业大学,2016.

[10] 李喜艳.奶牛乳腺上皮细胞中赖氨酸蛋氨酸配比模式对酪蛋白合成的影响及机理研究[D].硕士学位论文.北京:中国农业科学院,2011.

[11] 张航.不同日粮模式及长链脂肪酸对奶牛乳腺乳脂合成的影响及其机理研究[D].博士学位论文.呼和浩特:内蒙古农业大学,2014.

[12] ROBINSON T L,SUTHERLAND I A,SUTHERLAND J.Validation of candidate bovine reference genes for use with real-time PCR[J].Veterinary Immunology and Immunopathology,2007,115(1/2):160-165.

[13] 宗灿华.奶牛乳腺主要乳成分合成代谢的转录组学研究[D].博士学位论文.哈尔滨:东北农业大学,2013.

[14] O'BOYLE N J,CONTRERAS G A,MATTMILLER S A,et al.Changes in glucose transporter expression in monocytes of periparturient dairy cows[J].Journal of Dairy Science,2012,95(10):5709-5719.

[15] THREADGOLD L C,KUHN N J.Glucose-6-phosphate hydrolysis by lactating rat mammary gland[J].International Journal of Biochemistry,1979,10(8):683-685.

[16] 赵柯.奶牛乳腺上皮细胞葡萄糖摄取的调控及其对乳成分合成的影响研究[D].博士学位论文.杭州:浙江大学,2011.

[17] ONG L,JIN Y,SONG I C,et al.2-(¹⁸F)-2-deoxy-D-glucose (FDG) uptake in human tumor cells is related to the expression of GLUT-1 and hexokinase Ⅱ[J].Acta Radiologica,2008,49(10):1145-1153.

[18] FUEGER P T,BRACY D P,MALABANAN C M,et al.Hexokinase Ⅱ overexpression improves exercise-stimulated but not insulin-stimulated muscle glucose uptake in high-fat-fed C57BL/6J mice[J].Diabetes,2004,53(2):306-314.

[19] 赵艳丽.乙酸与亮氨酸、乙酸与蛋氨酸互作效应对奶牛乳腺上皮细胞内乳成分合成的影响及其机理研究[D].博士学位论文.呼和浩特:内蒙古农业大学,2016.

[20] RAMAKRISHNAN B,QASBA P K.Structure-based design of beta 1,4-galactosyltransferase Ⅰ (β 4GAL-T1) with equally efficient N-acetylgalactosaminyltransferase activity:point mutation broadens β 4GAL-T1 donor specificity[J].The Journal of Biological Chemistry,2002,277(23):20833-20839.

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